Hum. Reprod. Advance Access published online on May 22, 2009
Human Reproduction, doi:10.1093/humrep/dep120
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Mitochondrial membrane potential disruption pattern in human sperm
1 Centro de Biotecnología en Reproducción, Universidad de La Frontera, Temuco, Chile 2 Department of Dermatology/Andrology Unit, University of Leipzig, Leipzig, Germany 3 Departamento de Medicina Interna, Facultad de Medicina, Universidad de La Frontera, Montevideo 870, Casilla 54-D, Temuco, Chile
4 Correspondence address. E-mail: jvillega{at}ufro.cl
BACKGROUND: Loss of mitochondrial membrane potential (
m) in spermatozoa is correlated with high levels of reactive oxygen species in semen, abnormal spermiogram parameters, and low success rates of IVF. In somatic cells, the loss of 
m is primarily associated with several mechanisms of cell death, mainly the activation of caspases. The impact of mitochondrial dysfunction on sperm function is still not fully elucidated, although disruption of 
m and activation of caspases are processes thoroughly studied in human ejaculates. Disruption of 
m in sperm can be externally triggered by the antineoplastic agent betulinic acid (BA). In this study, we determined whether caspase activation is necessary for the BA-induced disruption of 
m in human sperm.
METHODS: Viable and highly motile sperm cells were selected through a swim-up process and incubated with 90 µg/ml BA. To elucidate the caspase dependency of BA-triggered disruption of 
m, we used the pan-caspase inhibitor zVAD-fmk and the caspase-3/7 inhibitor DEVD-cho.
RESULTS: Exposing highly motile sperm to BA caused a specific disruption of 
m (P < 0.001 versus control) and a corresponding increase in caspase-3/7 activity (P < 0.001 versus control). Pre-incubation of the sperm with zVAD-fmk or DEVD-cho only partially inhibited BA-induced loss of 
m (P < 0.05 versus control).
CONCLUSION: We found that caspases directly participate in the loss of 
m caused by BA in human sperm cells. However, caspase-independent pathways may also be present.
Key words: human spermatozoa/mitochondrial membrane potential/caspases/betulinic acid
Submitted on September 30, 2008; resubmitted on March 26, 2009; accepted on April 9, 2009.